Acetylcholinesterase (AChE) staining is associated with terminal fields of the glossopharyngeal and chorda tympani nerves in the nucleus of the solitary tract (NST). To address AChE function at these sites, the location of the staining was examined at the fine structural level in combination with the labeling of chorda tympani nerve fibers with biotinylated dextran in golden Syrian hamsters. AChE staining was located in the endoplasmic reticulum of geniculate ganglion neuronal somata, and extracellularly, surrounding labeled chorda tympani terminal fibers and boutons in the NST. Neuronal profiles adjacent to these labeled fibers were stained less intensely, whereas most non-adjacent profiles were unstained. The location of staining is consistent with the secretion of AChE into the extracellular space by primary afferent chorda tympani fibers. AChE staining was reduced in the dextran-labeled chorda tympani fibers and terminals as well as adjacent non-labeled profiles 2 weeks following nerve transection and dextran application. The distribution of staining outside synapses and the loss of staining following denervation is suggestive of a non-cholinergic role for AChE in the intact gustatory system.